Search

Rice (Oryza sativa L.) is a major food crop worldwide. Plant height and yield are important agronomic traits of rice. Several genes regulating plant height and/or yield have been cloned. However, the molecular mechanisms coordinating plant height and yield are not fully characterized. Here, we report a novel gene, OstMAPKKK5 that encodes a truncated variant of a mitogen-activated protein kinase kinase kinase 5 (OsMAPKKK5) lacking an intact kinase domain. Transgenic plants overexpressing OstMAPKKK5 in indica cultivar 9311 showed increased plant height, grain length, grain width, 1000-grain weight, grain number per main panicle, and yield. Real-time quantitative PCR showed that OstMAPKKK5 was widely expressed in various tissues and developmental stages. The increased plant height and yield were attributed to enlarged cell size. Overexpression of OstMAPKKK5 led to higher contents of various forms of endogenous gibberellin (GA), especially the most common active forms, GA1, GA3, GA4. We concluded that OstMAPKKK5 positively regulates plant height and yield in rice by affecting cell size, and that its underlying mechanism is based on increased endogenous GA content. Keywords: OstMAPKKK5, Plant height and yield, Cell size, Gibberellin, Oryza sativa

Heading date is one of the most important traits in rice and regulated by multiple genes. Common wild rice is the ancestor of Asian cultivated rice and harbours abundant genetic diversity. To use wild rice resource in rice breeding, a set of 154 introgression lines (ILs) covering 93% of the genome of Jinghong common wild rice was constructed in the background 'Teqing', using 208 simple sequence repeat markers evenly distributed on 12 chromosomes. Among the ILs, the line JIL64 displayed late heading independent of photoperiod. Genetic analysis using the two F₂ populations crossed ''Teqing'/JIL64 and JIL64/'Teqing' revealed that late flowering was controlled by a recessive gene on chromosome 8 (designated early heading date 8, ehd8), and ehd8 was fine mapped to the 50‐kb region flanked by markers RM22221 and 64Indel4. Sequencing and qRT‐PCR demonstrated that LOC_Os08g01410 and LOC_Os08g01420 were deleted in JIL64 and may be associated with the late heading of Jinghong common wild rice. These findings lay a practical foundation for characterizing ehd8, and the ILs help to mine genes from Jinghong common wild rice.

Grain number per panicle is a major component of rice yield that is typically controlled by many quantitative trait loci (QTLs). The identification of genes controlling grain number per panicle in rice would be valuable for the breeding of high-yielding rice. The Oryza glaberrima chromosome segment substitution line 9IL188 had significantly smaller panicles compared with the recurrent parent 9311. QTL analysis in an F2 population derived from a cross between 9IL188 and 9311 revealed that qgnp7(t), a major QTL located on the short arm of chromosome 7, was responsible for this phenotypic variation. Fine mapping was conducted using a large F3 population containing 2250 individuals that were derived from the F2 heterozygous plants. Additionally, plant height, panicle length, and grain number per panicle of the key F4 recombinant families were examined. Through two-step substitution mapping, qgnp7(t) was finally localized to a 41 kb interval in which eight annotated genes were identified according to available sequence annotation databases. Phenotypic evaluation of near isogenic lines (NIL-qgnp7 and NIL-qGNP7) indicated that qgnp7(t) has pleiotropic effects on rice plant architecture and panicle structure. In addition, yield estimation of NILs indicated that qGNP7(t) derived from 9311 is the favorable allele. Our results provide a foundation for isolating qgnp7(t). Markers flanking this QTL will be a useful tool for the marker-assisted selection of favorable alleles in O. glaberrima improvement programs.

Environmental stresses influence the growth of plants and the productivity of crops. Salinity is one of the most important abiotic stresses for agricultural crops. Programmed cell death (PCD) is induced by a variety of biotic and abiotic stresses in plants, including high salinity treatment. OsPDCD5, an ortholog of the mammalian-programmed cell death 5 gene, is up-regulated under low temperature and salt treatments in rice (Oryza sativa). In this study, transgenic rice that constitutively expressed antisense-OsPDCD5 increased salt stress tolerance in unique ways by inhibiting PCD pathways and regulating specific groups of stress-related genes. Real-time quantitative PCR analysis showed that PCD pathways were inhibited under both normal conditions and salt stress in transgenic rice. Using transcript microarray analysis, 38 stress-related genes regulated in transgenic rice were identified, which appeared to render the transgenic rice adapted for salt stress conditions. Our results suggested that PCD pathways were inhibited and salt stress signaling pathways were regulated in transgenic rice even under normal growth conditions without stress. PCD was further inhibited and these stress-related genes were further regulated to increase tolerance to high salinity under salt stress conditions, thereby leading to build-up of stress protection mechanisms in transgenic rice.

With 4 figures and 1 table Abstract Oilseed rape (Brassica napus L.) is the second most important oilseed crop in the world, and greater production of rape oil is required to satisfy the growing demand for it. We previously found that the cell size and yield of rice (Oryza sativa) and cotton can be increased by over-expression of the RNA-binding domains (RRMs) of the flowering control locus A (FCA) protein. We also found that B. napus FCA-RRM2 shares a high level of sequence similarity with the RRM2 domains of other plants. We isolated B. napus FCA-RRM2 from variety No.1 ‘Nannongyou’ of Brassica napus. Transformation experiments demonstrated that over-expression of B. napus FCA-RRM2 increases plant size, organ size, cell size, plant productivity and oil content in transgenic rape plants by down-regulating the cell-cycle-related gene cyclin-B2-1, a known activator of cyclin-dependent kinase 1 at the G2/M boundary. These results provide a practical approach for the genetic improvement of B. napus and possibly provide a model for investigating cell size regulation.

Many rice breeding programs have currently reached yield plateaus as a result of limited genetic variability in parental strains. Dongxiang common wild rice (Oryza rufipogon Griff.) is the progenitor of cultivated rice (Oryza sativa L.) and serves as an important gene pool for the genetic improvement of rice cultivars. In this study, heterotic loci (HLs) associated with six yield-related traits were identified in wild and cultivated rice and investigated using a set of 265 introgression lines (ILs) of O. rufipogon Griff. in the background of the Indica high-yielding cultivar Guichao 2 (O. sativa L.). Forty-two HLs were detected by a single point analysis of mid-parent heterosis values from test cross F(1) offspring, and 30 (71.5%) of these HLs showed significantly positive effects, consistent with the superiority shown by the F(1) test cross population in the six yield-related traits under study. Genetic mapping of hsp11, a locus responsible for the number of spikelets per panicle, confirmed the utility of these HLs. The results indicate that favorable HLs capable of improving agronomic traits are available. The identification of HLs between wild rice and cultivated rice could lead to a new strategy for the application of heterosis in rice breeding.

Panicle-related traits are important agronomic traits which directly associated with grain yield. In this study, we investigated quantitative trait loci (QTLs) associated with panicle-related traits using a set of 265 introgression lines (ILs) of common wild rice (Oryza rufipogon Griff.) in the background of Indica high-yielding cultivar Guichao 2 (O. sativa L.). A total of 39 QTLs associated with panicle-related traits including panicle length (PL), primary branch number (PBN), secondary branch number (SBN), spikelet number per panicle (SPP) and spikelet density (SD), were detected in the ILs with single-point analysis. The alleles of 20 QTLs derived from wild rice showed positive effects, and some QTLs, such as, QPl1b for PL, QPbn8 for PBN, QSd4 and QSd11b for SD and QSpp4 for SPP showed larger positive effects, providing good candidates and useful information for marker-aided improvement of yield potential of rice. Most of the QTLs controlling SPP, SBN and SD were located in cluster or closely linked on chromosomes, and the directions of their additive effects were consistent, which explained the genetic basis of significant correlations between their phenotypic characters.

In rice (Oryza sativa L.), the number of panicles, spikelets per panicle and grain weight are important components of grain yield. These characteristics are controlled by quantitative trait loci (QTLs) and are derived from variation inherent in crops. As a result of the complex genetic basis of these traits, only a few genes involved in their control have been cloned and characterized. We have previously map-cloned a gene cluster including eight leucine-rich repeat receptor-like kinase (LRK) genes in Dongxiang wild rice (Oryza rufipogon Griff.), which increased the grain yield by 16%. In the present study, we characterized the LRK1 gene, which was contained in the donor parent (Dongxiang wild rice) genome and absent from the recurrent parent genome (Guichao2, Oryza sativa L. ssp. indica). Our data showed that rice LRK1 is a plasma membrane protein expressed constitutively in leaves, young panicles, roots and culms. The over-expression of rice LRK1 results in increased panicles, spikelets per panicle, weight per grain and enhanced cellular proliferation, leading to a 27.09% increase in total grain yield per plant. The increased number of panicles and spikelets per panicle are associated with increased branch number. Our data suggest that rice LRK1 regulates rice branch number by enhancing cellular proliferation. The functional characterization of rice LRK1 facilitates an understanding of the mechanisms involved in cereal crop yield, and may have utility in improving grain yield in cereal crops.

A simple protocol of transformation of cotton (Gossypium hirsutum L.) at a high frequency has been developed via Agrobacterium mediation, coupled with the use of embryogenic calli as explants. Embryogenic calli derived from only one to two somatic embryogenic calli lines of two Chinese cotton cultivars, the cvs. Ekang 9 and Jihe 321 which have low embryogenic potency were first inoculated with the A. tumefaciens strain LBA4404 harbouring binary vector pBin438 carrying a synthetic Bacillus thuringiensis-active CrylAc and API-B chimeric gene. Infected embryogenic calli were co-cultivated for 48 h and were then moved on to the selection medium with kanamycin (100 mg/l) for 7-8 weeks. Then, the kanamycin-resistant calli (Km r ) subcultured in proliferation medium would re-differentiate to form somatic embryos in 30 days. Cotyledon embryos were transferred to 100-ml Erlenmeyer flasks for germination and regeneration. Putative transformants were confirmed by polymerase chain reaction and Southern blot analysis. Forty-five regenerated plants were successfully transferred to soil, of which 12 proved to have the active Cry1Ac and API-B chimeric gene. Insect resistance was tested by bioassay. The transgenic plants were highly resistant to cotton bollworm (Heliothis armigera) larvae, with mortality (insect resistance) ranging from 95.8 to 100%. In comparison with the methods used in Agrobacterium-mediated transformation of cotton hypocotyls or cotyledons, about 6 months are saved by using the method presented in this paper to obtain a large number of transgenic plants.

Common wild rice (Oryza rufipogon Griff.) is an important genetic reservoir for rice improvement. We investigated a quantitative trait locus (QTL), qGP5-1, which is related to plant height, leaf size and panicle architecture, using a set of introgression lines of O. rufipogon in the background of the Indica cultivar Guichao2 (Oryza sativa L.). We cloned and characterized qGP5-1 and confirmed that the newly identified gene OsEBS (enhancing biomass and spikelet number) increased plant height, leaf size and spikelet number per panicle, leading to an increase in total grain yield per plant. Our results showed that the increased size of vegetative organs in OsEBS-expressed plants was enormously caused by increasing cell number. Sequence alignment showed that OsEBS protein contains a region with high similarity to the N-terminal conserved ATPase domain of Hsp70, but it lacks the C-terminal regions of the peptide-binding domain and the C-terminal lid. More results indicated that OsEBS gene did not have typical characteristics of Hsp70 in this study. Furthermore, Arabidopsis (Arabidopsis thaliana) transformed with OsEBS showed a similar phenotype to OsEBS-transgenic rice, indicating a conserved function of OsEBS among plant species. Together, we report the cloning and characterization of OsEBS, a new QTL that controls rice biomass and spikelet number, through map-based cloning, and it may have utility in improving grain yield in rice.